基于Nrf2调控氧化应激研究麦味养肺汤对肺纤维化小鼠的保护作用及机制  

Protective effect and mechanism of Maiwei Yangfei Decoction on pulmonary fibrosis mice based on Nrf2 regulation of oxidative stress

作　　者：魏云 王晶[1,2] 韩迪 黄同杏 柏乐 陈立炜 徐泳 周贤梅 WEI Yun;WANG Jing;HAN Di;HUANG Tong-xing;BAI Le;CHEN Li-wei;XU Yong;ZHOU Xian-mei(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;the First Clinical Medical College,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Chinese Medicine,School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学附属医院,江苏南京210029 [2]南京中医药大学第一临床医学院,江苏南京210023 [3]南京中医药大学中医学院·中西医结合学院,江苏南京210023

出　　处：《中国中药杂志》2023年第24期6682-6692,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82074358);江苏省研究生实践创新计划项目(SJCX23_0751);江苏省研究生科研创新计划项目(KYCX22_1947,KYCX23_2176);大学生创新创业训练国家级项目(202310315030Z)。

摘　　要：探讨麦味养肺汤(Maiwei Yangfei Decoction,MWYF)对肺纤维化(pulmonary fibrosis,PF)小鼠的作用及其机制。制备MWYF水煎液,超高效液相色谱-三重四极杆串联质谱法(UPLC-MS/MS)检测其主要成分。雄性C57BL/6J小鼠随机分为对照组、模型组以及MWYF低(MWYF-L)、中(MWYF-M)、高(MWYF-H)剂量组和吡非尼酮(PFD)组,每组10只。除对照组外,其余各组用博莱霉素(bleomycin,BLM)气管滴注法构建肺纤维化模型,次日起给予生理盐水或MWYF或PFD灌胃处理。每日观察小鼠饮水饮食、行动、毛发;苏木精-伊红(HE)染色、马松(Masson)染色和CT观察肺组织病理改变;碱水解法检测肺组织中羟脯氨酸(HYP)水平;免疫组织化学法观察Ⅲ型胶原(COL3)、纤连蛋白(fibronectin)表达情况;反转录实时荧光定量PCR(RT-qPCR)检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原α1(COL1α1)、COL3、波形蛋白(vimentin)mRNA表达水平;试剂盒检测肺组织和血清中氧化应激指标超氧化物歧化酶(SOD)、丙二醛(MDA);免疫荧光检测核因子E2相关因子2(Nrf2)蛋白入核情况;蛋白质免疫印迹法和RT-qPCR检测肺组织中Nrf2、过氧化氢酶(CAT)、血红素加氧酶1(HO-1)蛋白及mRNA表达水平。UPLC-MS/MS共检测出12种化学成分。动物实验显示,MWYF可以改善PF小鼠肺泡炎症、胶原沉积及纤维化,增加小鼠体质量,下调肺组织中HYP、α-SMA、COL1α1、COL3、fibronectin、vimentin等纤维化指标的表达。此外,MWYF可以提高PF小鼠肺组织和血清中SOD活力,上调Nrf2表达水平并促进其向细胞核内转移,上调下游抗氧化靶基因CAT、HO-1水平,进而减少脂质代谢产物MDA的积累。综上,MWYF能够显著改善PF小鼠肺组织病理损伤和纤维化,其机制可能与激活Nrf2通路调控氧化应激有关。This study explored the effect and mechanism of Maiwei Yangfei Decoction(MWYF)on pulmonary fibrosis(PF)mice.MWYF was prepared,and its main components were detected by ultra-high-performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-MS/MS).Male C57BL/6J mice were randomly divided into a control group,a model group,a pirfenidone(PFD)group,and low-,medium-,and high-dose MWYF groups,with 10 mice in each group.The PF model was induced in mice except for those in the control group by intratracheal instillation of bleomycin(BLM),and model mice were treated with saline or MWYF or PFD by gavage the next day.The water consumption,food intake,hair,and activity of mice were observed daily.The pathological changes in lung tissues were observed by hematoxylin-eosin(HE)staining,Masson staining,and CT scanning.The level of hydroxyproline(HYP)in lung tissues was detected by alkaline hydrolysis.Immunohistochemistry was used to observe the expression of collagen typeⅢ(COL3)and fibronectin.The mRNA expression levels ofα-smooth muscle actin(α-SMA),typeⅠcollagenα1(COL1α1),COL3,and vimentin were detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Superoxide dismutase(SOD)and malondialdehyde(MDA)kits were used to detect oxidative stress indicators in lung tissues and serum.The nuclear translocation of nuclear factor E2-related factor 2(Nrf2)protein was detected by immunofluorescence.The protein and mRNA expression levels of Nrf2,catalase(CAT),and heme oxygenase 1(HO-1)in lung tissues were detected by Western blot and RT-qPCR.Twelve chemical components were detected by UPLC-MS/MS.Animal experiments showed that MWYF could improve alveolar inflammation,collagen deposition,and fibrosis in PF mice,increase body weight of mice,and down-regulate the expression of fibrosis indexes such as HYP,α-SMA,COL1α1,COL3,fibronectin,and vimentin in lung tissues.In addition,MWYF could potentiate the activity of SOD in lung tissues and serum of PF mice,up-regulate the

关 键 词：肺纤维化 麦味养肺汤 氧化应激 NRF2 

分 类 号：R285.5[医药卫生—中药学]